体外培养蓖麻蚕蛹精巢组织细胞对核型多角体病毒的敏感性分析  

Susceptibility of the in vitro Cultured Cells from Spermary Tissue of Philosamia cynthia ricini to Nucleopolyhedroviruses

作　　者：王文兵[1] 倪爱民[1] 李兵[2] 张月华[3] 徐安英[3] 沈卫德[2] 

机构地区：[1]江苏大学生命科学研究院,江苏镇江212013 [2]苏州大学基础医学与生命科学学院,江苏苏州215123 [3]中国农业科学院蚕业研究所,江苏镇江212018

出　　处：《蚕业科学》2010年第2期354-358,共5页ACTA SERICOLOGICA SINICA

基　　金：国家重点基础研究发展计划"973"项目(No.2005CB12-1005);江苏省高技术研究(农业)项目(No.BG2007322)

摘　　要：为开展核型多角体病毒与宿主互作关系的研究,建立蓖麻蚕(Philosamia cynthia ricini)细胞体外扩增体系,分析其对不同核型多角体病毒的敏感性差异。采用剪切分离后的蓖麻蚕蛹精巢组织材料,将分散的细胞在TNM-FH培养基中经数月换液培养后,获得了扩增的蓖麻蚕蛹精巢细胞系(BMC1)。BMC1的倍增时间约为65h。病毒的感染性实验表明:BMC1细胞对苜蓿银纹夜蛾核型多角体病毒(AcNPV)比较敏感,在感染的细胞中可以明显地观察到多角体;家蚕核型多角体病毒(BmNPV)的多角体蛋白基因启动子可以在BMC1细胞中表达,但难以观察到多角体形成和细胞的病变,提示BMC1可以作为BmNPV的稳定化表达受体;而蓖麻蚕核型多角体病毒(PcrNPV)单独感染BMC1细胞时,无明显病变,只有在与AcNPV混合感染时可观察到PcrNPV多角体形成。BMC1细胞对不同核型多角体病毒的敏感性具有差异,不同核型多角体病毒的感染性差异的机制有待深入解析。To study the interactive relationship between nucleopolyhedroviruses （NPVs） and their hosts, the in vitro proliferation cell lines from Philosamia cynthia ricini pupal spermary tissue were established and their susceptibility to different nucleopolyhedroviruses were analyzed. By splitting the spermary tissue of Philosamia cynthia ricini pupa, the separated cells were cultured in TNM-FH medium. After several months of culture, the cell line （named as BMC1 ） turned into the stage of proliferation. The population doubling time of BMCl was about 65 hours. The results of viral infection test indicated that BMC1 cells were fairly sensitive to Autographa colifomica NPV （AcNPV）. The polyhedra could be observed in the infected BMCl cells easily. The promoter of polyhedrin gene of Bombyx mori NPV （BmNPV） could be expressed in BMC1 cells. However. formation of the Dolvhedra could not be found, and the infection svmotoms were not observed as well, suggesting that BMC1 cells might be used for stable expression of BmNPV. Philosamia cynthia ricini NPV （PcrNPV） alone could not cause obvious infection symptoms. Only a mixed infection with AcNPV could lead to the expression of late polyhedrin gene and formation of PcrNPV polyhedra. The mechanism that results in the difference of susceptibility of BMC1 cells to nucleopolyhedroviruses infection remains for further studies.

关 键 词：蓖麻蚕 细胞系 精巢 核型多角体病毒 感染 

分 类 号：S885.2[农业科学—特种经济动物饲养]